Genomic signatures of host adaptation in group B Salmonella enterica ST416/ST417 from harbour porpoises.

A type of monophasic group B Salmonella enterica with the antigenic formula 4,12:a:- ("Fulica-like") has been described as associated with harbour porpoises (Phocoena phocoena), most frequently recovered from lung samples. In the present study, lung tissue samples from 47 porpoises found along the Swedish coast or as bycatch in fishing nets were analysed, two of which were positive for S. enterica. Pneumonia due to the infection was considered the likely cause of death for one of the two animals. The recovered isolates were whole genome sequenced and found to belong to sequence type (ST) 416 and to be closely related to ST416/ST417 porpoise isolates from UK waters as determined by core-genome MLST. Serovars Bispebjerg, Fulica and Abortusequi were identified as distantly related to the porpoise isolates, but no close relatives from other host species were found. All ST416/417 isolates had extensive loss of function mutations in key Salmonella pathogenicity islands, but carried accessory genetic elements associated with extraintestinal infection such as iron uptake systems. Gene ontology and pathway analysis revealed reduced secondary metabolic capabilities and loss of function in terms of signalling and response to environmental cues, consistent with adaptation for the extraintestinal niche. A classification system based on machine learning identified ST416/417 as more invasive than classical gastrointestinal serovars. Genome analysis results are thus consistent with ST416/417 as a host-adapted and extraintestinal clonal population of S. enterica, which while found in porpoises without associated pathology can also cause severe opportunistic infections.

Individually tailored internet-based treatment for young adults and adults with panic attacks: randomized controlled trial.

BACKGROUND: Previous studies on Internet-based treatment with minimal therapist guidance have shown promising results for several specific diagnoses. OBJECTIVE: To (1) investigate the effects of a tailored, therapist-guided, Internet-based treatment for individuals with reoccurring panic attacks, and (2) to examine whether people in different age groups (18-30 years and 31-45 years) would respond differently to the treatment. METHODS: We recruited 149 participants from an online list of individuals having expressed an interest in Internet treatment. Screening consisted of online questionnaires followed by a telephone interview. A total of 57 participants were included after a semistructured diagnostic interview, and they were randomly assigned to an 8-week treatment program (n = 29) or to a control condition (n = 28). Treatment consisted of individually prescribed cognitive behavior therapy text modules in conjunction with online therapist guidance. The control group consisted of people on a waitlist who later received treatment. RESULTS: All dependent measures improved significantly immediately following treatment and at the 12-month follow-up. The between-group effect size on the primary outcome measure, the Panic Disorder Severity Scale, was d = 1.41 (95% confidence interval 0.81-1.95) at posttreatment. The within-group effect size from pretreatment to 12-month follow-up was d = 1.66 (95% confidence interval 1.14-2.35). Age group had no effect, suggesting that age did not influence the outcome. CONCLUSIONS: Tailoring an Internet-based treatment can be a feasible approach in the treatment of panic symptoms and comorbid anxiety and depressive symptoms. Younger adults benefit as much as adults over 30 years and up to 45 years of age. TRIAL REGISTRATION: Clinicaltrials.gov NCT01296321; http://www.clinicaltrials.gov/ct2/show/NCT01296321 (Archived by WebCite at http://www.webcitation.org/65wddsqlL).

Injured cyclists with focus on single-bicycle crashes and differences in injury severity in Sweden.

While cycling is promoted as a clean, energy-efficient mode of transport generating physical activity, the number of injured cyclists must decrease to achieve traffic safety goals. The extent of the single bicycle crashes (SBCs) and crash causes are rather well studied. This study expands this knowledge by focusing on differences in injury severity. The aim of the study is to investigate the relationship between injury severity and characteristics of the crash and the cyclist with focus on SBCs. Furthermore, injury risk is calculated for different age classes and sexes, as well as for different purposes of the trip. The results are based on injured cyclists in Sweden (N = 105,836) registered in STRADA, 2010-2019, by both the police and accident and emergency departments (A&Es), with a special focus on injury severity reported by the A&Es. Binary logistic regression was applied to analyse how the odds of being severely injured differed for different cyclists and situations. Results from of the National Travel Survey, 2011-2016, were used to study differences in distance travelled with respect to sex, age group and purpose of the trip. Given that the cyclist is injured in an SBC, the results show a higher probability of being severely injured (maximal AIS 3 or more) for cyclists 45 years or older compared to younger cyclists, for males compared to females and for cyclists not wearing a helmet compared to cyclists wearing a helmet. A higher probability for severe injury was also found for crashes occurring during leisure trips compared to work/school trips, crashes occurring during weekdays compared to weekends and crashes at intersections and road stretches compared to pedestrian and cycle paths. Furthermore, the risk of being severely injured in an SBC per km travelled was higher for cyclists aged 45 and older and during a leisure trip.

Detection of an IMI-2 carbapenemase-producing Enterobacter asburiae at a Swedish feed mill.

Occurrence of multidrug resistant Enterobacteriaceae in livestock is of concern as they can spread to humans. A potential introduction route for these bacteria to livestock could be animal feed. We therefore wanted to identify if Escherichia spp., Enterobacter spp., Klebsiella spp., or Raoutella spp. with transferable resistance to extended spectrum cephalosporins, carbapenems or colistin could be detected in the environment at feed mills in Sweden. A second aim was to compare detected isolates to previous described isolates from humans and animals in Sweden to establish relatedness which could indicate a potential transmission between sectors and feed mills as a source for antibiotic resistant bacteria. However, no isolates with transferable resistance to extended-cephalosporins or colistin could be identified, but one isolate belonging to the Enterobacter cloacae complex was shown to be carbapenem-resistant and showing carbapenemase-activity. Based on sequencing by both short-read Illumina and long-read Oxford Nanopore MinIon technologies it was shown that this isolate was an E. asburiae carrying a bla IMI-2 gene on a 216 Kbp plasmid, designated pSB89A/IMI-2, and contained the plasmid replicons IncFII, IncFIB, and a third replicon showing highest similarity to the IncFII(Yp). In addition, the plasmid contained genes for various functions such as plasmid segregation and stability, plasmid transfer and arsenical transport, but no additional antibiotic resistance genes. This isolate and the pSB89A/IMI-2 was compared to three human clinical isolates positive for bla IMI-2 available from the Swedish antibiotic monitoring program Swedres. It was shown that one of the human isolates carried a plasmid similar with regards to gene content to the pSB89A/IMI-2 except for the plasmid transfer system, but that the order of genes was different. The pSB89A/IMI-2 did however share the same transfer system as the bla IMI-2 carrying plasmids from the other two human isolates. The pSB89A/IMI-2 was also compared to previously published plasmids carrying bla IMI-2, but no identical plasmids could be identified. However, most shared part of the plasmid transfer system and DNA replication genes, and the bla IMI-2 gene was located next the transcription regulator imiR. The IS3-family insertion element downstream of imiR in the pSB89A was also related to the IS elements in other bla IMI-carrying plasmids.

Fto colocalizes with a satiety mediator oxytocin in the brain and upregulates oxytocin gene expression.

Single nucleotide polymorphisms in the fat mass and obesity-associated (FTO) gene have been associated with obesity in humans. Alterations in Fto expression in transgenic animals affect body weight, energy expenditure and food intake. Fto, a nuclear protein and proposed transcription co-factor, has been speculated to affect energy balance through a functional relationship with specific genes encoding feeding-related peptides. Herein, we employed double immunohistochemistry and showed that the majority of neurons synthesizing a satiety mediator, oxytocin, coexpress Fto in the brain of male and female mice. We then overexpressed Fto in a murine hypothalamic cell line and, using qPCR, detected a 50% increase in the level of oxytocin mRNA. Expression levels of several other feeding-related genes, including neuropeptide Y (NPY) and Agouti-related protein (AgRP), were unaffected by the FTO transfection. Addition of 10 and 100 nmol oxytocin to the cell culture medium did not affect Fto expression in hypothalamic cells. We conclude that Fto, a proposed transcription co-factor, influences expression of the gene encoding a satiety mediator, oxytocin.

Functional coupling analysis suggests link between the obesity gene FTO and the BDNF-NTRK2 signaling pathway.

BACKGROUND: The Fat mass and obesity gene (FTO) has been identified through genome wide association studies as an important genetic factor contributing to a higher body mass index (BMI). However, the molecular context in which this effect is mediated has yet to be determined. We investigated the potential molecular network for FTO by analyzing co-expression and protein-protein interaction databases, Coxpresdb and IntAct, as well as the functional coupling predicting multi-source database, FunCoup. Hypothalamic expression of FTO-linked genes defined with this bioinformatics approach was subsequently studied using quantitative real time-PCR in mouse feeding models known to affect FTO expression. RESULTS: We identified several candidate genes for functional coupling to FTO through database studies and selected nine for further study in animal models. We observed hypothalamic expression of Profilin 2 (Pfn2), cAMP-dependent protein kinase catalytic subunit beta (Prkacb), Brain derived neurotrophic factor (Bdnf), neurotrophic tyrosine kinase, receptor, type 2 (Ntrk2), Signal transducer and activator of transcription 3 (Stat3), and Btbd12 to be co-regulated in concert with Fto. Pfn2 and Prkacb have previously not been linked to feeding regulation. CONCLUSIONS: Gene expression studies validate several candidates generated through database studies of possible FTO-interactors. We speculate about a wider functional role for FTO in the context of current and recent findings, such as in extracellular ligand-induced neuronal plasticity via NTRK2/BDNF, possibly via interaction with the transcription factor CCAAT/enhancer binding protein ß (C/EBPß).

Functional impairment after treatment with pectoral muscle flaps because of deep sternal wound infection.

OBJECTIVE: Pectoral muscle flaps (PMF) are effective in terminating protracted sternal wound infections (SWI) but long-term outcome remains uncertain. Therefore, the aim of this study was to evaluate long-term outcome in patients treated with PMF. DESIGN: Thirty-four of 263 patients revised because of deep SWI from 1991--2005 were treated with PMF. Of the 21 patients alive, 11 had left-sided, two right-sided and eight bilateral procedures. Sternal debridement without closure of the sternum was done in 17 patients. Nineteen of 21 patients responded to a questionnaire. RESULTS: At follow-up on average 5.9 years (range 1.9--14.8 years) after surgery 63% (12/19) experienced unstable chest. Two thirds (12/18) reported problems carrying a grocery bag and 37% (7/19) had problems putting on a coat. Reduction of power and mobility was more common in the right arm and shoulder even in patients with left-sided PMF. Thirty-two percent (6/19) would have preferred alternative treatment if possible to avoid sternal instability even if healing had been substantially delayed. CONCLUSIONS: Surgery with PMF and sternal debridement was associated with long-term disability, which appeared to be significant in one third of the patients. The function of the right arm and shoulder was affected more often despite the majority of procedures being left-sided suggesting that loss of skeletal continuity of the chest wall is more disabling than loss of pectoral muscle function.

Low rate of COVID-19 seroconversion in health-care workers at a Department of Infectious Diseases in Sweden during the later phase of the first wave; a prospective longitudinal seroepidemiological study.

Background: Health-care workers are at risk of contracting and transmitting SARS-CoV-2. The aim of this study was to investigate the prevalence of SARS-CoV-2 IgG antibodies and the rate of seroconversion in an environment with high exposure to SARS-CoV-2. Methods: 131 health-care workers at the Department of Infectious Diseases in Västerås, Sweden, were included in the study. Abbott's SARS-COV-2 IgG immunoassay was used with a signal cut-off ratio of ≥1.4. Every third week from the beginning of May, blood samples were drawn, and the participants completed a questionnaire regarding symptoms consistent with COVID-19 and the result of any SARS-CoV-2 PCR performed since the last sampling occasion. Participants with IgG antibodies against SARS-CoV-2 were re-sampled only on the sixth and last occasion. Results: At the start of the study, 18 (15%) participants had SARS-CoV-2 IgG antibodies. At the end, 25 (19%) of 131 participants were seropositive. One case of asymptomatic infection was detected, and two cases with PCR-confirmed COVID-19 did not develop IgG antibodies. Conclusion: The low rate of seroconversion during the study suggests that it is possible to prevent transmission of SARS-COV-2 in a high-exposure environment. Compliance with adequate infection control guidelines is the likely explanation of our findings.

Evaluation of 11 SARS-CoV-2 antibody tests by using samples from patients with defined IgG antibody titers.

We evaluated the performance of 11 SARS-CoV-2 antibody tests using a reference set of heat-inactivated samples from 278 unexposed persons and 258 COVID-19 patients, some of whom contributed serial samples. The reference set included samples with a variation in SARS-CoV-2 IgG antibody titers, as determined by an in-house immunofluorescence assay (IFA). The five evaluated rapid diagnostic tests had a specificity of 99.0% and a sensitivity that ranged from 56.3 to 81.6% and decreased with low IFA IgG titers. The specificity was > 99% for five out of six platform-based tests, and when assessed using samples collected ≥ 22 days after symptom onset, two assays had a sensitivity of > 96%. These two assays also detected samples with low IFA titers more frequently than the other assays. In conclusion, the evaluated antibody tests showed a heterogeneity in their performances and only a few tests performed well with samples having low IFA IgG titers, an important aspect for diagnostics and epidemiological investigations.

Effects of Gender-confirming Pitch-raising Surgery in Transgender Women a Long-term Follow-up Study of Acoustic and Patient-reported Data.

OBJECTIVES: Transgender women often seek to feminize their voice so that it becomes congruent with their gender identity. Many receive voice therapy (VT) with good results. Some also need pitch-raising surgery. The purpose of this study was to investigate long-term effects and to compare outcomes between cricothyroid approximation (CTA) and glottoplasty (GP). METHODS: This study included retrospective data from 24 patients (35-67 years). Eleven patients had undergone CTA and 13 had undergone GP. Audio recordings were performed in a sound-treated booth and patients answered questionnaires before and after VT, post surgery, and at 1-year follow-up. Fundamental frequency (fo) measures were extracted from voice range profiles (VRPs) and speech range profiles (SRPs) and were compared with cisgender data. RESULTS: Minimum fo in VRP increased significantly only after CTA (95 to 123 Hz). Maximum fo in VRP was significantly lowered after GP (765 to 652 Hz), even more after CTA (677 to 475 Hz). Speaking mean fo increased significantly after VT and post surgery, and was thereafter stable (CTA 167 Hz, GP 169 Hz). The maximum fo in the SRP increased only after VT. The frequency ranges were strongly reduced after CTA. Patients were in general satisfied at follow-up and rated the GP outcomes more favorably than CTA. CONCLUSIONS: Both surgical methods have advantages and disadvantages. The very restricted speaking and physiological frequency ranges, which do not favor an optimal female voice, were mainly found after the irreversible CTA. Thus, CTA is no longer performed at our hospital, whereas surgical techniques based on GP are being developed further. We strongly recommend the use of VRPs to evaluate treatment effects.

An analysis of cyclists' speed at combined pedestrian and cycle paths.

Objective: In Sweden, cyclists, pedestrians, and moped riders share the space on combined pedestrian and cycle paths, and their speeds may differ greatly. Both actual speed and speed differences can potentially influence the number of accidents on the shared paths. As a starting point, this article studies the speed component and how cyclists' speed varies at pedestrian and cycle paths depending on the day, week, and year; road user composition; and road design.Methods: Three data sources were used: Existing measurements of cycle speed and flow in 3 different Swedish municipalities, Eskilstuna (1 site, January-December 2015), Linköping (6 sites, 4 weeks in September-October 2015), and Stockholm (10 sites, 1-5 days in August-September 2015); complementary measurements of cycle speed and flow in Linköping (4 sites, 1-10 days in August-September 2016) and Stockholm (1 site, only part of 2 days in August 2016) were also conducted within the project, in addition to roadside observations of bicycle types at the 5 new sites.Results: The average speed of cyclists on the paths varied between 12.5 and 26.5 km/h. As expected, the lower average speeds were found in uphill directions, near intersections, and on paths with high pedestrian flows. The higher speeds were found in downhill directions and on commuter routes. In all, 70%-95% of road users observed on pedestrian and cycle paths were cyclists, and 5%-30% were pedestrians. The most common type of bicycle was a comfort bike, followed by a trekking bike. Electric-assisted bicycles and racer bikes occurred at all sites, with proportions of 1%-10% and 1%-15%, respectively. The 2 sites with the highest proportion of electric-assisted bicycles and racer bicycles also had the highest average speeds. The differences in average speed throughout the day, week, and year could only be assessed at one of the sites. Only small differences were found, with the most noticeable being that the average speed was lower in January and February (13.8 km/h) compared to the rest of the year (15.3-16.1 km/h). The average speed was also lower during daytime (14.7 km/h) than during other parts of the day (15.4-15.8 km/h).Conclusions: The relationship between bicycle type and measured speed was not entirely clear, but the results suggest that paths with higher proportions of electric and racer bicycles have higher average speeds. There also appears to be a connection between average speed and the width of the distribution; that is, the higher the average speed, the wider the speed distribution. More research is needed on how speed levels and speed variance affect accident risk.

A SELDI-TOF MS study of the genetic and post-translational molecular heterogeneity of eosinophil cationic protein.

Eosinophil cationic protein (ECP), a secretory protein of the eosinophil granulocyte, is a basic and highly heterogeneous protein. This heterogeneity is dependent on polymorphisms in the ECP gene and post-translational modifications, and it affects the functional properties of the protein in terms of cytotoxicity. The aim of this study was to further investigate the molecular heterogeneity, hence, an affinity capture assay based on an antigen-antibody interaction with the surface-enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI-TOF MS) technique was developed. Of three monoclonal antibodies tested, that is, EG2, 614, and 652, the 614 mab was chosen for the experiments. ECP heterogeneity of single individuals was studied in extracts of purified blood eosinophils, and the presence of approximately 5 major molecular species was demonstrated in each subject. ECP from subjects with different ECP 434(G>C) genotypes (arg97thr) showed mass differences corresponding to the amino acid shift from arginine to threonine. ECP purified from pooled leukocytes of large numbers of healthy blood donors demonstrated an extensive mass heterogeneity with approximately 10 major molecular species. By the use of a variety of glucosidases it was shown that this heterogeneity was mainly due to N-linked oligosaccharides on which sialic acid, galactose, and acetylglucosamine was positioned. We conclude that the SELDI-TOF MS technique using specific monoclonal antibodies is a convenient and versatile tool; by means of this technique, we could detect both genetic and post-translational causes of the molecular heterogeneity of the eosinophil cationic protein.

The 434(G>C) polymorphism within the coding sequence of Eosinophil Cationic Protein (ECP) correlates with the natural course of Schistosoma mansoni infection.

Schistosomiasis is a chronic parasitic infection with over 200 million people infected worldwide. In Schistosoma mansoni infections, parasite-derived eggs get trapped in the liver, causing the formation of granulomas, which may develop into periportal fibrosis and portal hypertension, and thus severe morbidity. Eosinophil cationic protein (ECP) is a secretory protein of eosinophil granulocytes that efficiently kills the larval stage of S. mansoni, but also affects fibroblast functions. We have investigated the prevalence of the ECP gene polymorphism 434(G>C) in two African populations, from an S. mansoni endemic area in Uganda (n=297) and from a non-endemic area in Sudan (n=78), and also compared these with a Swedish population (n=209). The genotype frequencies in the Ugandan population differed significantly from both the Sudanese and Swedish populations (P<0.001). In the Ugandan population there was a significant association between genotype and prevalence of infection (P=0.03), with lower prevalence in subjects with the GG genotype compared with GC (P=0.02) and CC (P=0.03). There was also a trend towards an association with periportal fibrosis (P=0.08) in the Ugandan population. This suggested association was confirmed when the predominant tribe (n=212) was analysed separately (P=0.004). Our results suggest that ECP may be an important protein, both in the immune response against S. mansoni and in the development of periportal fibrosis. The results also suggest genetic selection towards the ECP 434CC genotype in populations living in S. mansoni endemic areas.